AIM: To investigate the expression of matrix metalloproteinases (MMPs), membrane type MMPs (MT-MMPs) and their inhibitors (TIMPs) in human primary cultured prostatic cells and malignant prostate cell lines.METHODS: Reverse transcription-polymerase chain reaction-based measurements of the mRNA levels of MMP-2, MMP-7, MMP-9, MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 in relation to the house-keeping gene glyceraldehydes phosphate dehydrogenase were performed in cancerous and non-cancerous prostatic tissue samples, in primary cell cultures of epithelial cells, in both fibroblasts and smooth-muscle cells as stromal cells, and in the human malignant prostatic cell lines DU-145, LNCaP and PC-3.RESULTS: MMP-2 was mainly expressed in stromal cells. MMP-7 and MMP-9 showed their highest values in epithelial cells. MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 were found both in stromal and in epithelial cells, but there were some differences between the expression in fibroblasts and smooth-muscle cells. Different expression was also observed between the cells deriving from the primary cell cultures, the benign cell line BPH-1, and the malignant cell lines LNCaP, DU-145, and PC-3.CONCLUSION: These results concerning different expression of MMPs and TIMPs in cells from prostatic tissue suggest that a better insight into changes observed in prostatic tissue needs studies on cells cultured from the tissue.

Asthma ; diagnosis ; Child ; Humans ; Perception

OBJECTIVETo investigate the effect of hypoxia on the human pulmonary artery smooth muscle cells two pore domain potassium channels TASK-1 and the regulation of non-receptor tyrosine kinase c-Src in this process.

METHODSThe cultured human pulmonary artery smooth muscle cells (hPASMCs) were divided into: normal group, hypoxia 30 minute group, hypoxia 6 hours group and hypoxia 48 hour group, and hypoxia 48 hour + PP2 group, hypoxia 48 hour + PP3 group, hypoxia 48 hour + bpV group. Flow cytometry was used to analyze the cell cycle, RT-PCR and Western blot technique were carried out to detect the expression changes of TASK-1 mRNA and protein in different groups.

RESULTS(1) Cell Cycle Show: Compared with normal control group, with prolonged hypoxia, the percentages of hPASMCs in S phases of cell cycle were increased. While compared with hypoxia 48 hour group, the percentages of hypoxia 48 hour + PP2 group hPASMCs in S phases of cell cycle were decreased. The expression of TASK-1 mRNA on hPASMCs in acute hypoxia 6 hour group was increased, while the expression of TASK-1 protein on hPASMCs in the acute and chronic hypoxia group was decreased, and the expression of TASK-1 mRNA on hPASMCs in the chronic hypoxia group was decreased; After pre-incubation of a potent and selective inhibitor of the Src family of protein tyrosine kinases PP2, the expression of TASK-1 mRNA and protein in hypoxia 48 hour group was increased, however after pre-incubation of the inhibitor of the Src family of protein tyrosine phosphatase bpV, the expression of TASK-1 protein in hypoxia 48 hour group was decreased.

CONCLUSIONHypoxia promotes human pulmonary artery smooth muscle cell proliferation, and non-receptor tyrosine kinase c-Src may participate in the expression of two pore domain potassium channels TASK-1 regulated by hypoxia. Therefore, we hypothesized that TASK-1 channels and c-Src participatein the acute and chronic hypoxic human pulmonary vasoconstriction.

Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology ; Nerve Tissue Proteins ; metabolism ; Potassium Channels, Tandem Pore Domain ; metabolism ; Pulmonary Artery ; cytology ; RNA, Messenger ; Vasoconstriction ; src-Family Kinases ; metabolism

OBJECTIVEPrepare konjac glucomannan-hydroxypropyl methyl cellulose (HPMC) compression coated tablets and study the effects of the formulation, technics and in vitro dissolution condition on drug release behavior to elevate the colon-specific effects of preparation.

METHODBerberine hydrochloride core tablets were prepared by wet granulation technique and konjac glucomannan-HPMC mixture as the coating layer were used with compression coated technique. The effects of the formulation and technics on drug release behavior were investigated by dissolution test. The erosion of coat layer during dissolution test was investigated.

RESULTDrug almost not released in dissolution medium stimulating gastric and intestinal condition, and released completely by coating layer erosion and rupture by enzyme in stimulating colonic condition. Drug release decreased with decreasing the ratio of konjac glucomannan-HPMC and increasing coat weight (P < 0.05), compression force was not found to be a significant factor on drug release. Drug release increased with increasing the concentration of beta-mannase in dissolution medium (P < 0.05), rotation speed has no effect on drug release. The release of drug was correlative with erosion of coat layer. The mechanism of drug release were diffusion and erosion.

CONCLUSIONThe konjac glucomannan-HPMC compression coated tablets was a promising delivery system for drugs to be delivered to the colon.

Administration, Oral ; Amorphophallus ; chemistry ; Berberine ; administration & dosage ; chemistry ; pharmacokinetics ; Colon ; metabolism ; Drug Compounding ; methods ; Drug Delivery Systems ; Hypromellose Derivatives ; Mannans ; chemistry ; isolation & purification ; Methylcellulose ; analogs & derivatives ; chemistry ; Plants, Medicinal ; chemistry ; Tablets, Enteric-Coated

Objective To investigate homografting vascular endothelial progenitor cells(EPCs)for preventing restenosis formation of carotid artery in New Zealand white rabbit models.Methods EPCs of New Zealand white rabbits were isolated,confirmed and expanded though the injured carotid arterial endothelium of rabbit model induced by dilatation with a 2.5 F balloon;and then EPCs were transplanted into the injured endothelium of the cells transplantation group(n=13,3 of them were transplanted with fluorencently-labeled- EPCs),while equal volume of saline without EPCs was injected into the injured endothelium in the control group(n=8).Histopathology was performed at 4 days after transplantation for the 2 rabbits,with fluorencently-labeled-EPCs.All of the rest remained rabbits were killed 4 weeks later for histological examinations.Results The histopathological slides showed that the fluorescence-positive expression existed in the injured endothelium 4 days after transplantation.At 4 weeks after the EPCs transplantation,there were less restenosis and less vascular wall thickening in the rabbits of cells transplantation group than those of the control group(P＜0.01).Conclusion The local interventional homografting heterogeneous endothelial progenitor cells can prevent restenosis after the carotid artery angioplasty in New Zealand White rabbit model. (J Intervent Radiol,2007,16:95-98)

BACKGROUNDActinomycosis is a rare indolent infectious disease caused by Actinomyces. Although pulmonary actinomycosis is thought to be more prevalent in developing countries, data from developing countries are scanty. This study was to reveal the current situation of pulmonary actinomycosis in developing countries and the difference from that in developed countries.

METHODSPatients fulfilling the inclusion criteria for pulmonary actinomycosis from Peking Union Medical College Hospital in China between January 2003 and December 2014 were retrospectively analyzed. Baseline characteristics, clinical symptoms, underlying diseases, diagnostic methods, pulmonary function test results, chest computed tomography (CT) tests, fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) tests, initial diagnosis, treatment and prognosis were retrieved from medical records and analyzed.

RESULTSTwenty-six patients were included in this study (mean age 52.0 + 13.1 years). The ratio of male to female was 1.17:1. Most common clinical symptoms were cough (15/26), sputum (12/26) and hemoptysis (12/26). Chest CT findings presented as masses (13/26), nodules (10/26) and infiltrates (3/26). FDG-PET had an increased standardized uptake value and 4/6 patients were misdiagnosed as malignancy. Many kinds of antibiotics were used in the treatment of pulmonary actonomycosis and all got favorable results. Five patients receiving complete resection of the lesion were cured without postoperative use of antibiotic.

CONCLUSIONSPulmonary actinomycosis is a rare disease even in developing countries, and both misdiagnosis and missed diagnosis are common. FDG-PET seems useless in the differential diagnosis, and complete resection of the pulmonary lesion without postoperative antibiotic therapy might be enough to achieve cure.

Actinomycosis ; diagnosis ; diagnostic imaging ; metabolism ; Adult ; Aged ; China ; Female ; Fluorodeoxyglucose F18 ; Humans ; Lung Diseases ; diagnosis ; diagnostic imaging ; metabolism ; Male ; Middle Aged ; Positron-Emission Tomography ; Radiography ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo find sensitive and specific micro-metastic markers for prostate cancer.

METHODSUsing nested reverse transcription-PCR, we examined the expression of PSA, hK2 and PSMA mRNA in peripheral blood mononuclear cells of 51 patients with prostate cancer, 33 patients with benign prostate hyperplasia (BPH) and 32 normal young people.

RESULTSThe expression rates of PSA, hK2 and PSMA mRNA were 52.9%, 43.1% and 64.7%, respectively in prostate cancer group, and 6.2%, 7.7% and 4.6%, respectively in control group (BPH patients and normal young people) with statistical significance (P < 0.01). Although the expression rate of PSA and hK2 mRNA increased with cancer progression, there was no statistical significance among patients in different stages. The expression rate of PSMA mRNA was higher than that of PSA and hK2 mRNA in each clinical stage.

CONCLUSIONPSMA mRNA expression detected by nested RT-PCR is of greater value for the diagnosis, therapy choice and prognostic evaluation of prostate cancer patients.

Aged ; Antigens, Surface ; blood ; Biomarkers, Tumor ; blood ; Glutamate Carboxypeptidase II ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; blood ; pathology ; Prostatic Neoplasms ; blood ; pathology ; Tissue Kallikreins ; blood

OBJECTIVETo explored the relationship between genotype of Bruton's tyrosine kinase (BTK) gene and X-linked agammaglobulinemia (XLA).

METHODSSix patients who were clinically suspected as XLA based on immunological results were studied. Peripheral blood samples were collected for DNA extraction. The 19 exons and exon-intron boundaries of the BTK gene were amplified by PCR, and the products were directly sequenced.

RESULTSAll of the 6 patients were confirmed to have XLA due to the mutations in exons of the BTK gene. Among these, 3 mutations were located in the kinase domain (TK), 2 were located in pleckstrin homology (PH) domain, and 1 was located in Src homology (SH2) domain. The mutations have included 3 missense mutations, i.e., c.1105C to T (p.L369F), c.82C to T(p.R28C) and c.1754T to C (p.V585A), 2 nonsense mutations, i.e., c.1834C to T (p.Q612X) and c.37C to T (p.R13X). One patient was found to have complex (missense and nonsense) mutations, i.e., c.1802-1803TT to GC (p.F601C) and c.1803-1804insC (p.T602fsX603). There were 3 novel mutations (p.F601C, p.T602fsX603 and p.V585A). The mothers of 5 patients were also detected with BTK gene mutations, among whom 4 were demonstrated to be carriers and one was normal (her son had p.V585A mutation). Therefore, p.V585A was a de novo mutation.

CONCLUSIONDetection of BTK gene mutation can confirm clinical diagnosis which is critical for patients to take regular immunoglobulin replacement therapy for life. Early genetic diagnosis can also identify carriers and make genetic counseling possible.

Agammaglobulinemia ; enzymology ; genetics ; Base Sequence ; Child ; Child, Preschool ; Codon, Nonsense ; Female ; Genetic Diseases, X-Linked ; enzymology ; genetics ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense ; Protein-Tyrosine Kinases ; genetics

OBJECTIVETo review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.

DATA SOURCESThe data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.

STUDY SELECTIONMainly original articles and critical reviews written by major pioneer investigators of this field were selected.

RESULTSThe research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.

CONCLUSIONST4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.

Bacterial Proteins ; metabolism ; DNA-Binding Proteins ; Gene Transfer, Horizontal ; Helicobacter pylori ; genetics ; metabolism ; pathogenicity ; Multigene Family

OBJECTIVETo access the feasibility of employing metabonomics method in clinical studies. This pilot study intends to introduce nuclear magnetic resonance (NMR)-based metabonomics method to elucidate the metabolism of non-syndromic cleft lip and/or palate (NSCLP) patients.

METHODSHigh-resolution 1H NMR spectroscopy was performed on blood plasma obtained from NSCLP and non-malformed children. All signal of 1H NMR spectra were recognized within MESTRE-v4.7, and the 1H NMR spectra integration into bins (or buckets) across the spectral regions of bin 0.04 was performed automatically in MESTRE-v4.7. The resulting data matrix was further analyzed, which was performed by SIMCA-P 11.0. The principal component analysis (PCA) was applied to the centered data to explore any clustering behavior of the samples.

RESULTSThe results demonstrated the metabonomic difference in plasma between NSCLP and non-malformed children at least lies in 3-Hydroxybutyrate gamma-CH3, arginine and valine. Arginine excretion appeared to be higher in the non-malformed children population, while NSCLP population excreted higher concentrations of 3-Hydroxybutyrate gamma-CH3 and valine.

CONCLUSIONThe present study clearly demonstrated the great potential of the NMR-based metabonomics approach in elucidating the NSCLP plasma metabolism and the possibility of application in clinic diagnosis and screening.

Child ; Cleft Lip ; Cleft Palate ; Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; Pilot Projects